US8895054B2 - Methods for joint lubrication and cartilage wear prevention making use of glycerophospholipids - Google Patents
Methods for joint lubrication and cartilage wear prevention making use of glycerophospholipids Download PDFInfo
- Publication number
- US8895054B2 US8895054B2 US12/411,855 US41185509A US8895054B2 US 8895054 B2 US8895054 B2 US 8895054B2 US 41185509 A US41185509 A US 41185509A US 8895054 B2 US8895054 B2 US 8895054B2
- Authority
- US
- United States
- Prior art keywords
- dmpc
- liposomes
- joint
- cartilage
- dppc
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active, expires
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/66—Phosphorus compounds
- A61K31/683—Diesters of a phosphorus acid with two hydroxy compounds, e.g. phosphatidylinositols
- A61K31/685—Diesters of a phosphorus acid with two hydroxy compounds, e.g. phosphatidylinositols one of the hydroxy compounds having nitrogen atoms, e.g. phosphatidylserine, lecithin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/06—Antipsoriatics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P41/00—Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
- A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/24—Materials or treatment for tissue regeneration for joint reconstruction
Definitions
- This invention generally concerns liposomes and methods for their therapeutic use.
- Joint dysfunctions affect a very large portion of the population. Sufficient biolubrication is a prerequisite for proper joint mobility, which is crucial for prevention and amelioration of degradative changes of the joint 1 .
- osteoarthritis A common joint dysfunction is osteoarthritis, with prevalence exceeding 20 million in the United States alone 2 .
- the etiology of osteoarthritis is multifactorial, including inflammatory, metabolic and mechanical causes 3-5 .
- risk factors involved are age, gender, obesity, occupation, trauma, atheromatous vascular disease and immobilization 1, 3-7 .
- osteoarthritis may arise as a result of articular cartilage breakdown; or conversely, subchondral bone sclerosis may actually precede cartilage degeneration and loss 8, 9 . Once articular cartilage is injured, damage progresses 10 .
- Articular cartilage forms a smooth, tough, elastic and flexible surface that facilitates bone movement.
- the synovial space is filled with the highly viscous synovial fluid (SF), containing hyaluronic acid (HA) and the glycoprotein lubricin 12-14 .
- HA is a polymer of D-glucuronic acid and D-N-acetylglucosamine, which is highly unstable and degrades under the inflammatory conditions of osteoarthritis 15, 16 .
- Lubricin is composed of ⁇ 44% proteins, ⁇ 45% carbohydrates and ⁇ 11% phospholipids (PL) 12-14 , of which ⁇ 41% are phosphatidylcholines (PCs), ⁇ 27% phosphatidylethanolamines (PE) and ⁇ 32% sphingomyelins 17-19 . These PL are referred to as “surface-active phospholipids” (SAPL).
- SAPL surface-active phospholipids
- the PE and PC of SAPL contain two hydrocarbon chains, one of which is the monounsaturated oleic acid (18:1).
- Boundary lubrication in which layers of lubricant molecules separate opposing surfaces, occurs under loading of articular joints 17, 18, 20 .
- HA was thought to be the major lubricant 21 , however, a recent tribiological study states that HA “by itself . . . is not responsible for the nearly frictionless boundary biolubrication found in articular cartilage”, but may contribute to load bearing and wear protection 22 .
- Many reports have shown lubricin to play the major role in the lubricating properties of synovial fluid 12, 14, 19, 20, 23, 24 .
- Pickard et al 25 and Schwartz and Hills 19 demonstrated that phospholipids defined as surface active phospholipids (SAPL) of lubricin facilitate joint lubrication in articular cartilage.
- SAPL surface active phospholipids
- DPPC in the form of multilamellar vesicles (MLV) has a phase transition temperature in which solid ordered (SO) to liquid disordered (LD) phase transition occurs of 41.4° C.
- U.S. Pat. No. 6,800,298 discloses dextran-based hydrogel compositions containing lipids, particularly phospholipids, for lubrication of mammalian joints.
- Klein et al. 31 summarized various issues of joint lubrication at the molecular level. They point to the potential role of highly-hydrated brush-like charged macromolecules at the surface of cartilage as major contributors to cartilage lubrication 31-33 .
- the present invention is based on the discovery of a liposomal platform for joint lubrication and on studies of the effect of different phospholipids (PL) compositions, liposomal sizes and lamellarity on joint friction and/or on cartilage wear, using a cartilage-on-cartilage apparatus that mimics articular joints.
- PL phospholipids
- a novel formulation based on a liposome system comprising PL is proposed, for introduction into synovial joints in order to improve or restore joint mobility. It was been found that the novel liposomal platform is effective as a lubricant as well as for reducing cartilage wear.
- liposomes comprising one or more membranes with at least one phospholipid (PL) of the group consisting of a glycerophospholipid (GPL) having two, being the same or different, C 12 -C 16 hydrocarbon chain and a sphingolipid (SPL) having a C 12 -C 18 hydrocarbon chain, the one or more membranes having a phase transition temperature in which solid ordered (SO) to liquid disordered (LD) phase transition occurs, the phase transition temperature being within a temperature of about 20° C. to about 39° C., the use being for lubrication and/or reducing wear rate of joints (cartilage) having a joint temperature which is above the phase transition temperature of the membrane.
- PL phospholipid
- GPL glycerophospholipid
- SPL sphingolipid
- a pharmaceutical composition for joint lubrication and/or reducing wear rate of joints having a joint temperature and comprising a physiologically acceptable carrier and liposomes; the liposomes comprising one or more membranes with at least one phospholipid (PL) of the group consisting of glycerophospholipid (GPL) having two, being the same or different, C 12 -C 16 hydrocarbon chains and a sphingolipid (SPL) having a C 12 -C 18 hydrocarbon chain; the one or more membranes having a phase transition temperature in which solid ordered (SO) to liquid disordered (LD) phase transition occurs, the phase transition temperature being within a temperature of about 20° C. to about 39° C. and being below said joint temperature.
- PL phospholipid
- GPL glycerophospholipid
- SPL sphingolipid
- the GPL, SPL or their combination form liposomes, preferably liposomes with a mean diameter greater than about 0.3 ⁇ m, typically greater than about 0.5 ⁇ m and at times greater than about 0.8 ⁇ m.
- the mean diameter of the liposomes is usually less than about 10 ⁇ m, typically less than about 8, 7, 6 or 5 ⁇ m and at times less than 3.5 ⁇ m.
- the liposomes may be a single-membrane liposome or may be, according to one embodiment, multilamellar vesicles (MLV) liposomes. According to other embodiments the liposomes may also be large multivesicular vesicles (LMVV) or dehydrated rehydrated vesicles (DRV) liposomes.
- LMVV large multivesicular vesicles
- DUV dehydrated rehydrated vesicles
- said C 12 -C 16 or C 12 -C 18 hydrophobic chains are saturated.
- FIG. 1 is a bar graph showing the friction coefficients (static and dynamic) obtained for various lubricating media, including inflamed synovial fluid (ISF); histidine buffer (HB, 5 mM), dispersions comprising multilamellar vesicles (MLV, carried in 5 mM HB, the lipids being at a concentration range of between 35-140 mM) with the phospholipid being DMPC, MLV comprising DMPC, or DMPC-cholesterol, or mixture of DMPC and 2000 PEG-DSPE DMPC or a mixture of DMPC and DPPC, or small unilamellar vesicles (SUV) comprising DMPC. All measurements were performed at 37° C. under contact pressure of 2.4 MPa (30N load) and sliding velocity of 1 mm/s. Saline was used as a control.
- ISF inflamed synovial fluid
- HB histidine buffer
- MLV multilamellar vesicles
- FIG. 2 is a graph showing cartilage wear as a function of sliding distance in the presence of two different potential carrier media HB and saline, the cartilage wear being measured by analyzing the glycosamineglycan (GAG) content weight in the debris present in the aqueous medium of the cartilage as a result of wearing.
- GAG glycosamineglycan
- FIG. 3 is a graph showing cartilage wear as a function of sliding distance in the presence of DOPC, HSPC, DPPC and DMPC, all dispersed in HB.
- FIG. 4 is a graph showing cartilage wear as a function of sliding distance in the presence of DPPC, DMPC and a mixture of DMPC/DPPC, all dispersed in HB.
- FIG. 5 is a graph showing cartilage wear as a function of sliding distance in the presence of ISF, ISF+HA and ISF+ DMPC/DPPC.
- FIG. 6 is a bar graph showing the constant wear rates obtained with different lubricating fluids (Saline, HB or HA) and phospholipids (DOPC, HSPC, DPPC, DMPC/DPPC or DMPC), used as liposome additives, following a run-in period.
- lubricating fluids Saline, HB or HA
- DOPC phospholipids
- FIG. 7 is a graph showing the effect of the various lubricants and media on total phospholipid concentration, in cartilage specimens from healthy individuals after being subjected to similar friction tests in the presence of the different lubricants. The controls were not subjected to friction tests.
- FIG. 8 is a graph showing PC concentration as a function of vertical depth into cartilage where cartilage specimens were subjected to similar friction tests in the presence of: DMPC-MLV (0.8-3.5 ⁇ m in diameter) 141 mM in 5 mM HB ( ⁇ ); DMPC-SUV ( ⁇ 100 nm in diameter) 141 mM in 5 mM HB ( ⁇ ); or HB alone 5 mM (x); sliced into discs and tested for their DMPC concentration as a function of cartilage depth.
- FIGS. 9A-9F are scanning electron microscope (SEM) micrographs of cartilage specimens in the presence and absence of lubricating media and friction tests.
- FIG. 9A is a micrograph of healthy cartilage, showing its naturally occurring lipidic vesicle structures on the surface (x3000);
- FIG. 9B is a micrograph of arthritic cartilage (x3000); and healthy cartilage subjected to friction tests in the presence of the following lubricants: saline (x6000, FIG. 9C ); ISF (x800 FIG. 9D ); DMPC-SUV (x800, FIG. 9E ); and DMPC-MLV (x6000 FIG. 9F ).
- phospholipids selected from glycerophospholipids (GPL) and sphingolipids (SPL), are potential substituents for the naturally-occurring lipidic globular structures, being capable of reducing friction and protecting against cartilage wear.
- the GPL is carrying a phosphocholine headgroup (phosphatidylcholine, PC-based lipid) or a phosphoglycerol headgroup (phosphatidylglycerol, PG-based lipid), and the SPL is a ceramide (N-acyl sphingosine carrying a phosphocholine headgroup, also referred to as N-acyl sphigosyl-phosphocholine (SM-based lipid).
- N-acyl sphingosine carrying a phosphocholine headgroup also referred to as N-acyl sphigosyl-phosphocholine (SM-based lipid).
- the PG is negatively charged over broad pH range as evident from it negative zeta potential.
- the hydrophobic part of the PC and PG includes 2 hydrocarbon (e.g. acyls and alkyls) chains.
- the SM also has two hydrophobic hydrocarbon chains of which one is the chain of the sphingoid base itself and the other is N-acyl chain.
- PC, SM and PG in which the hydrocarbon chains is above 12 carbon atoms are all cylinder like in shape as their packing parameter is in the range of 0.74-1.0. They form lipid bilayers which above the SO to LD phase transition become highly hydrated and vesiculate to form lipid vesicles (liposomes) 34, 35 .
- the PC and PG liposome bilayer can be either in a solid ordered (SO) phase (previously referred to as gel or solid phase), or in a liquid disordered (LD) phase (previously referred to as liquid crystalline or fluid phase) 34 .
- SO solid ordered
- LD liquid disordered
- the transformation between the SO to LD phases involves an endothermic, first order phase transition referred to as the main phase transition.
- T m is the temperature in which the maximum change in the heat capacity change during the SO to LD phase transition occurs.
- T m and the temperature range of the SO to LD phase transition of PCs depend, inter alia, on PC hydrocarbon chain composition. In the LD phase (but not in the SO phase), the charged phopshocholine and phosphoglycerol head group are highly hydrated.
- PGs and SM have T m that are similar to that of the corresponding PC (the same length of substituting hydrocarbon chain(s)).
- T m of DMPG is identical to that of DMPC, namely, 23° C.
- DPPG or N-palmitoyl SM is identical to that of DPPC, namely, 41.4° C.
- the PL in accordance with the invention may also be a PG- or SM-based lipid.
- a mixture of two or more PLs may be used, as long as the mixture formed is in a LD state and the lipid headgroups are highly hydrated, when in situ (either at the articular region of a healthy or a dysfunctioning joint).
- liposomal systems for joint lubrication which are chemically stable, oxidative-damage-resistant and free of HA.
- a liposome comprising at least one PL selected from glycerophospholipid (GPL) or sphingolipid (SPL), for joint lubrication is provided.
- GPL glycerophospholipid
- SPL sphingolipid
- a liposome comprising at least one PL selected from glycerophospholipid (GPL) or sphingolipid (SPL), for the preparation of a pharmaceutical composition for joint lubrication.
- GPL glycerophospholipid
- SPL sphingolipid
- the liposomes in accordance with both aspects being characterized in that they comprise one or more membranes with at least one phospholipid (PL) of the group consisting of a glycerophospholipid (GPL) having two, being the same or different, C 12 -C 16 hydrocarbon chains and a sphingolipid (SPL) having a C 12 -C 18 hydrocarbon chain.
- PL phospholipid
- GPL glycerophospholipid
- SPL sphingolipid
- the phase transition temperature in which solid ordered (SO) to liquid disordered (LD) phase transition occurs is within a temperature range of about 20° C. to about 39° C.
- SO solid ordered
- LD liquid disordered
- the liposomes are used to lubricate joints that have a joint temperature that is somewhat higher than the phase transition temperature. Accordingly the liposomes are in an LD phase within the joint.
- the fact that the joint temperature is typically only slightly (e.g. within the range of about 1° C. to about 15
- said C 12 -C 16 or C 12 -C 18 hydrophobic chains are saturated.
- the above conditions are cumulative, namely, the selection of PL (either a single PL or a combination of PL with additional PLs) contained in the liposome is so that the liposome will have SO-LD phase transition temperature between about 20° C. to about 39° C.
- the liposomal systems making use the said GPL or SPL further encompass one or more of the following, all of which require to exhibit a liposomal system having a phase transition temperature as defined herein:
- the GPL or SPL have alkyl, alkenyl or acyl C 12 to C 16 hydrocarbon chain.
- the two chains may be the same or different.
- One particular embodiment concerns the use of liposomes having GPL or SPL with at least one C 14 acyl chain.
- Another particular embodiment concerns the use of a GPL having C 14 and C 16 acyl chains.
- Another particular embodiment concerns the use of liposomes having SPL with a C 16 acyl chain.
- Another particular embodiment concerns the use of a combination of any of the above liposomes.
- GPL or SPL have a ionic headgroup and, according to embodiments of the invention, this headgroup is highly ionized at a wide range of pH.
- a wide range may be defined by a pH between 3 and 14.
- the GPL as well as the SPL are highly hydrated, namely, the number of water molecules per lipid headgroup is at least about 6; 7 or at times at least 8 water molecules that are complexed to the ionized head group of the GPL or SPL.
- the GPL or SPL are capable of forming MLV (as well as the other type of liposomes mentioned above), preferably MLV having a mean diameter above 0.3 ⁇ m.
- the MLV are defined by a mean diameter in the range of between 0.3 ⁇ m and 5 ⁇ m.
- the MLV are defined by a mean diameter in the range of between 0.8 ⁇ m and 3.5 ⁇ m.
- the MLV or the other types of liposomes that may be used in accordance with the invention should not include in their bilayers a membrane active sterol, such as cholesterol.
- a membrane active sterol is defined as affecting short- and long-range lipid order within membranes, minimizing volume, and decreasing membrane permeability.
- the sterol should possess 1), a flat, fused ring system, 2), a hydroxyl or other small polar group at position 3, 3), a “cholesterol-like” tail, and 4), a small area per molecule ( ⁇ 40 ⁇ 2 when assembled at the air/water interface at a surface pressure of 12 mN/m).
- compositions of the invention preferably do not contain propylene glycol.
- compositions of the invention preferably do not contain dextran.
- a particular group of GPLs encompassed by one or more of the above embodiments comprise a GPL carrying a phosphocholine headgroup (PC or SM-based lipids).
- PC phosphocholine headgroup
- SM-based lipids phosphocholine headgroup
- One preferred PC in accordance with the invention is dimyristoylphosphatidylcholine (DMPC).
- Non-limiting examples of PC-based lipids which may be used in accordance with the invention comprise 1,2-dipalmitoyl-sn-glycero-3-phosphocoline (DPPC, T m 41.4° C.); 1,2-dipentadecanoyl-sn-glycero-3-phosphocoline (C15, T m 33.0° C.), albeit, these two being suitable only when combined with one or more other lipids so as to form a liposomal system having a phase transition temperature as defined herein.
- SPL which may be in accordance with the invention comprise a sphingomyelin (SM) carrying a phosphocholine headgroup, and non-limiting examples include N-palmitoyl SM T m 41.0° C.
- SM sphingomyelin
- T m values of various PC-based lipids may be found in “Thermotropic Phase Transitions of Pure Lipids in Model Membranes and Their Modifications by Membrane Proteins”, John R. Silvius, Lipid - Protein Interactions , John Wiley & Sons, Inc., New York, 1982, and also in the Lipid Thermotropic Phase Transition Data Base—LIPIDAT, and in Marsh (1990) 36 .
- the MLV liposomes (or the other liposomes useful according to the invention) have an offset temperature (upper limit) of the SO to LD phase transition which is not higher than 15° C. from the temperature in situ, i.e. in the joint, within the range of about 20° C. to about 39° C.
- the MLV liposomes are formed from GPL, SPL or their combination, and the SO to LD phase transition temperature described above thus concerns MLV liposomes which are formed from GPL, SPL and combinations thereof, thus providing a liposome in which the PLs or their mixture are in LD phase.
- a particular embodiment in accordance with the invention concerns the use of DMPC-MLV or DMPC/DPPC-MLV for the preparation of a replacement of naturally-occurring cartilage PL, namely as a cartilage lubricant and wear reducer.
- MLV have major practical advantages as well. They can be prepared simply and at low cost. DMPC and DPPC are both resistant to oxidative damage and stable for long periods of time. Furthermore, these PCs are already approved for human use.
- the mole ratio between DMPC and DPPC depends on the temperature of the joint to be treated and is designed such that the T m of the combination provides MLV in LD phase.
- a suitable ratio is about 0.6/1.0 which provides MLV in LD phase at a joint temperature between 35° C. to 39° C.
- a method for lubricating a joint of a mammal comprises administering into a cavity of said joint containing synovial fluid an amount of liposomes effective to yield a lubricating effect.
- the GPL or the mixture thereof with additional PLs be in a LD phase, in situ, at the joint region to be lubricated therewith.
- the method of the invention may be used to treat, alleviate, retard, prevent, manage or cure any articular disorder or symptoms arising there from which is associated with joint dysfunction.
- articular disorder shall be held to mean any affliction (congenital, autoimmune or otherwise), injury or disease of the articular region which causes degeneration, pain, reduction in mobility, inflammation or physiological disruption and dysfunction of joints.
- the disorder may be associated with reduced joint secretion and lubrication as well as from complications of knee and hip replacement.
- the joint in accordance with the invention may be any one of the knee, hip, ankle, shoulder, elbow, tarsal, carpal, interphalangeal and intervertebral.
- Specific articular disorders include, but are not limited to, deficiencies of joint secretion and/or lubrication arising from arthritis, including conditions of joint erosion in rheumatoid arthritis, osteoarthritis, osteoarthritis in rheumatoid arthritis patients, traumatic joint injury (including sports injury), locked joint (such as in temporomandibular joint (TMJ)), status post arthrocentesis, arthroscopic surgery, open joint surgery, joint (e.g. knee or hip replacement) in mammals, preferably humans.
- a preferred disorder to be treated or prevented by the method of the invention is osteoarthritis.
- the method of the present invention could be used as a prophylactic measure to prevent future damage or degeneration.
- the PL based MLV liposomes could be administered intra-articularly to athletes intermittently throughout their career to minimize the risk of stress related injury or cartilage degeneration.
- the method of the present invention may be used exclusive of, or as an adjunct to, anti-inflammatory agents, analgesic agents, muscle relaxants, anti-depressants, or agents that promote joint lubrication commonly used to treat disorders associated with joint stiffness, such as arthritis.
- a combined therapeutic approach is beneficial in reducing side effects associated with agents, such as non-steroidal, anti-inflammatory drugs (NSAIDs), commonly used to prevent, manage, or treat disorders such as osteoarthritis associated with reduced joint lubrication.
- NSAIDs non-steroidal, anti-inflammatory drugs
- a combined therapeutic approach may also be advantageous in increasing efficacy of treatment.
- the administration of the liposomes into an articular cavity of a patient may be by a method chosen from the group consisting of intra-articular injection, arthroscopic administration or surgical administration.
- the invention also provides, in accordance with yet another aspect of the invention, a pharmaceutical composition for joint lubrication comprising a physiologically acceptable carrier and liposomes comprising at least one PL selected from GPL or SPL as defined herein.
- the physiologically acceptable carrier is hyaluronic acid (HA) or histidine buffer (HB).
- the composition may also include polymers such as those described by Klein et. al. 31 .
- composition according to the invention is preferably in a form suitable for administration by a route selected from intra-articular injection, arthroscopic administration or surgical administration.
- the amount of liposomes in the composition will vary depending on the liposome's PL composition, the disease, its severity and treatment regimen, as well a on the age, weight, etc., of the mammal to be treated.
- the amount for purposes herein is determined by such considerations as may be known in the art.
- the amount must be effective to achieve an improvement in the lubrication of the treated joint, namely, to reduce friction between the cartilages forming the joint, the improvement may be exhibited by clinical tests as well as by an improvement in the well-being of the subject undergoing said treatment (e.g. reduced pain in the afflicted joint, improvement in mobility).
- the effective amount is typically determined in appropriately designed clinical trials (dose range studies) and the person versed in the art will know how to properly conduct such trials in order to determine the effective amount.
- Table 1 Lipids used in this study and their sources are described in Table 1; all are >98% pure. Table 1 also presents the solid-ordered (SO) to liquid-disordered (LD) phase transition temperatures, T m , of phospholipid bilayers, 34-36 as well as the bilayer state at 37° C.
- SO solid-ordered
- LD liquid-disordered
- Phase transition Phase at temperature Lipid Chemical name (source) 37° C. (T m ), ° C. HSPC hydrogenated soybean SO 52.5 phosphatidylcholine (Lipoid, Ludwigshafen, Germany) DPPC 1,2-dipalmitoyl-sn-glycero-3- SO 41.4 phosphocholine (Avanti, Alabaster, AL, USA) DMPC 1,2-dimyristoyl-sn-glycero-3- LD 23.2 phosphocholine (Lipoid or Avanti) DOPC 1,2-dioleoyl-sn-glycero-3- LD ⁇ 21 phosphocholine) (Lipoid or Avanti) Mixture of LD 34 DMPC/ DPPC (0.6/0.1) Water: Water was purified using a WaterPro PS HPLC/Ultrafilter Hybrid system (Labconco, Kansas City, Mo.), providing pyrogen-free water with low levels of total carbons and inorgan
- Liposomes Multilamellar liposomes (MLV) were prepared by dissolving the desired lipids in tert-butanol, followed by lyophilization to form a dry “cake”. This was hydrated in low ionic strength (5 mM) histidine buffer (HB) pH 6.7, at a temperature at least 5° C. above the T m 34 .
- MLV were downsized to form small unilamellar vesicles ( ⁇ 100 nm, SUV) by stepwise extrusion through polycarbonate membranes (GE-Osmonics, Minnetonka, Minn.), starting with a 400-nm and ending with a 50-nm-pore-size membrane, using a 10-mL extrusion system (Northern Lipids, Vancouver, Canada) heated at least 5° C. above the T m 37 .
- polycarbonate membranes GE-Osmonics, Minnetonka, Minn.
- DMPC liposomes acted as the best friction reducers ( FIGS. 1 and 3 ). Therefore, DMPC-based liposomes were further investigated comparing liposomes composed of either DMPC alone, of different sizes and lamellarities, or of a DMPC/DPPC mixture (0.6:1.0 mole ratio), or of DMPC combined with cholesterol (2:1 mole ratio), or of DMPC combined with the lipopolymer mPEG-DSPE (95:5 mole ratio).
- the mPEG-DSPE used consists of a 2000 Dalton polyethylene glycol attached to the primary amino group of distearoyl phosphatidylethanolamine.
- Liposome characterization Liposomes were characterized for:
- PL concentration was also quantified as a function of cartilage depth. For this, cartilage specimens were sectioned by microtome into slices 20 or 50 ⁇ m thick, from the cartilage surface inwards, parallel to the face of the cartilage. PL concentration of each slice was quantified, after PL were extracted as mentioned above, by the modified Bartlett procedure 37, 38 .
- the reciprocating sliding amplitude was set to 1 mm, assuring full contact between the two cartilage samples during the wear test. Each test was carried out for 7.5 hours with an average reciprocating velocity of 4 m/min. This resulted in a maximum of 450,000 reciprocating cycles for each test, which amounts to a total sliding distance of 1800 m. Normal load of 60 N corresponding to contact pressure of 4.8 MPa, which is well in the range of physiological pressures in joints, was used. In order to compensate for any possible loss of cartilage components (which can occur, for example, from loose ends at the circumferential cylindrical cut surface of the plugs during agitation in the lubricating fluid), experiments with identical test conditions but with no load and no contact between the cartilage specimens were also conducted. The actual wear was calculated by subtracting the results at no load from these obtained when load was applied.
- a pair of frozen specimens was thawed, and the upper and lower specimens were fixed to a wear test loading mechanism and a bath, respectively.
- a volume of 1.5 ml of the lubricating fluid was placed in the bath and the reciprocating sliding motion started after applying a normal load (compensation at 0 N, and wear test at 60 N).
- glycosaminoglycans (GAGs) in articular cartilage is sulphated (mainly chondroitin sulfate and keratan sulfate) 61
- analyzing for GAG content 62 in the aqueous cartilage medium containing the wear particles was used to assess the wear of the two cartilage specimens.
- Cartilage structure Cartilage structure was examined by SEM. Specimens were preserved by rapid cooling in liquid nitrogen and kept under vacuum ( ⁇ 15 mbar) for 48 h to remove excess water. Next, specimens were mounted on stubs and sputter-coated with gold in a Polaron E5100 Sputter Coater (Watford, England). The specimens were examined using an FEI Quanta 200 scanning electron microscopy system (Polaron) using an accelerating voltage of 30 kV. Results Liposome carrier: it was found that the lubrication efficiency of HB is better than that of saline, or of ISF ( FIG. 1 ).
- liposomes dispersed in HB were better lubricants than liposomes dispersed in saline ( FIG. 2 ).
- liposome additives to be screened for their cartilage-lubricating abilities were all dispersed in HB.
- SAPL surface-active phospholipids tested were phosphatidylcholines (PCs), which are also naturally present in cartilage and synovial fluid.
- FIG. 4 shows the amount of cartilage wear found in the presence of DPPC, DMPC and a mixture of DMPC/DPPC, all dispersed in HB, as a function of sliding distance.
- Using liposomes composed of a mixture of close to ideally miscible PC's (no phase separation) may enable fitting their T m to suit a wide range of temperatures.
- the ratio of DMPC/DPPC can be adjusted, so that phase transition would take place at all physiological temperatures occurring in different conditions of osteoarthritis.
- ISF, ISF+DMPC/DPPC, ISF+HA In order to determine whether an addition of liposomes in ISF can reduce wear, comparative tests were performed in the presence of ISF alone and ISF+DMPC/DPPC. Of all the liposomes tested earlier, the mixture of DMPC/DPPC was chosen as it was found to be the most efficient lubricant additive (see FIGS. 3 and 4 ).
- ISF+DMPC/DPPC 150 mM was prepared by adding a mixture of DMPC/DPPC in HB with a concentration of 300 mM to ISF with a ratio of 1:1.
- FIG. 5 shows the amount of cartilage wear found in the presence of ISF, ISF+HA and ISF+DMPC/DPPC as a function of sliding distance. Similar behavior of wear to that in the presence of media alone and other liposome based lubricants was shown, where a relatively short run-in period was followed by a long constant wear rate period. Comparing cartilage wear in the presence of the different lubricants shows that although the addition of HA to ISF induces less wear than ISF alone, the addition of DMPC/DPPC mixture to ISF is much more effective in terms of reducing cartilage wear.
- FIG. 6 A summary of the constant wear rates following the run-in period, that were obtained with all the different lubricating fluids and additives is presented in FIG. 6 .
- the constant wear rate of cartilage in the presence of HB is lower in comparison with saline and therefore HB was selected as the preferred carrying media for all the tested liposome types. Screening the different liposomes shows that the lowest constant wear rate is achieved in the presence of the mixture of DMPC/DPPC.
- adding DMPC/DPPC to ISF reduces the constant wear rate by ⁇ 40% compared with ISF alone, while adding HA to ISF reduces the constant wear rate only by ⁇ 10%. Based on these results it was suggested that intra-articular injections of DMPC/DPPC may be used to improve cartilage lubrication in osteoarthritis patients.
- Friction and wear in cartilage lubricated with several DMPC-based liposomes were compared to that of ⁇ 100-nm unilamellar DMPC liposomes (DMPC-SUV).
- DMPC-MLV multilamellar DMPC liposomes
- DMPC-SUV ⁇ 100-nm unilamellar DMPC liposomes
- the efficacy as cartilage lubricants of DMPC-MLV enriched with lipids which are non-liposome-forming, although are common liposome components, such as cholesterol or mPEG-DSPE was studied.
- Cholesterol having a packing parameter of ⁇ 1.2 39 , was added at ⁇ 33 mole % to form DMPC/cholesterol-MLV, thus causing the transformation of the lipid bilayer from the solid-ordered (SO, if PL are below the T m ) or liquid-disordered (LD, if PL are above T m ) phase to a new physical phase termed liquid-ordered (LO) 43, 44 .
- SO solid-ordered
- LD liquid-disordered
- LO liquid-ordered
- DMPC-MLV Another component added to DMPC-MLV was the lipopolymer mPEG-DSPE, having a relatively low packing parameter of ⁇ 0.5 39 , which introduces a highly-hydrated extended steric barrier that surrounds the liposome 39, 45 .
- mPEG-DSPE was added at 5 mole % to form DMPC/mPEG-DSPE-MLV.
- the static and dynamic friction coefficients of DMPC-MLV in HB were lower than those obtained with DMPC/cholesterol-MLV in HB (0.040 and 0.036, respectively) or DMPC/mPEG-DSPE-MLV in H13 (0.022 and 0.023, respectively), as shown in FIG. 1 , and were similar to the low friction coefficients which exist in healthy synovial joints 46 .
- the static and dynamic friction coefficients of cartilage lubricated with DMPC-MLV were lower than those of cartilage lubricated with DMPC-SUV (0.045 and 0.036, respectively) which were only slightly lower than those of HB alone (0.053 and 0.037, respectively), FIG. 1 .
- K The partial specific adiabatic compressibility, is a measure of both the physical phase of the lipid bilayer (SO, LD or LO) and its hydration state, which is postulated herein to have an important contribution to the liposomes' efficacy as friction and wear reducers 45
- Values of K for DMPC, DPPC and hydrogenated soy phosphatidylcholine (HSPC) determined at 37° C. were 50.7, 31.2 and 33.3 ⁇ 10 ⁇ 6 mL/(g-atm), respectively.
- K values reflect the higher phase transition temperatures, T m , of DPPC and HSPC (41.4° C., 52.5° C.) than that of DMPC (23.2° C.) and thus the superiority of liposomes having a membrane having a phase transition in the range defined in the present invention (T m between 20° C. to 39° C. inclusive).
- T m phase transition temperatures
- DMPC/cholesterol liposomes (2:1 mole ratio K is reduced to 42.2 and 45.5 ⁇ 10 ⁇ 6 mL/(g-atm) at 24° C. and 37° C., respectively.
- PL levels in lubricated cartilage specimens The total PL (which includes naturally-occurring SAPLs and PLs from liposomes) levels of healthy cartilage specimens (thickness ⁇ 1200 ⁇ m), before and after being subjected to friction tests, in the presence of different lubricants and media, was measured. It can be seen ( FIG. 8 ) that the total PL concentration in cartilage lubricated with DMPC-MLV is the highest among all specimens tested.
- the PL concentration of cartilage obtained from healthy subjects and lubricated with FIB is higher than that of similar cartilage lubricated with saline or ISF, the latter (ISF), has similar PL levels to that of cartilage obtained from osteoarthritis patients.
- PC concentration as a function of cartilage depth (0-800 ⁇ m, in 20-50- ⁇ m increments) was measured after friction tests for specimens lubricated with DMPC-MLV and DMPC-SUV, both dispersed in HB, and for specimens lubricated with HB alone (control).
- cartilage lubricated with DMPC-MLV had the highest PC concentration near the cartilage surface ( FIG. 8 ).
- PC concentration reached a maximum at a depth of ⁇ 100 ⁇ m, below which, it decreased.
- FIG. 9A shows healthy cartilage, where naturally-occurring globular lipidic structures are dispersed on its porous surface, as previously shown on the surface of rat cartilage by Ohno et al. 28, 48 .
- the surface of osteoarthritic cartilage lacks these structures ( FIG. 9B ), as does friction-tested healthy cartilage lubricated with saline ( FIG. 9C ) or ISF ( FIG. 9D ), indicating poor protection against wear by these lubricants.
- FIG. 9E On the surface of cartilage lubricated with DMPC-SUV ( FIG. 9E ), very few lipidic structures can be noticed after friction testing.
- DMPC-MLV FIG. 9F ), large lipidic structures, resembling those on healthy cartilage, are present after friction testing.
- the present example was conducted to assess local reactions (histopathology of fermorotibial joint) at different times post intraarticular injection of DMPC-based MLV composed of either DMPC alone or of a DMPC/DPPC mixture (0.6:1.0 mole ratio) in Sprague Dawley (SD) rats.
- Animals 46 male SD rats aged 9 weeks old (purchased from Harlan Laboratories Ltd. Israel) were randomly assigned to 5 groups, 9 animals per group. The rats were maintained at Assaf Harofe Medical Center animal facility. The rats treated by intraarticular injection according to an initial assignment (Table 2, Group composition, treatments and Identification of animals). Rat in general good condition were included. All rats were treated on Day 1, and one group was treated again on Day 14. Both knees were treated; each knee was injected with 100 ⁇ L of one of the test liposomes or control substance. Cages were marked with a clear label and permanent marker. The rats were marked on their tail.
- the physiological solution that was used is 0/9% w/v sodium chloride (purchased from B. Braun batch No. 7281C12 exp. 08.2010).
- Table 3 summarizes the weights and treatment of each rat.
Abstract
Description
- Hills, B. A. Phospholipid and propylene glycol based lubricant. U.S. Pat. No. 6,133,249, 1998.
- Hills, B. A. Lubricant Composition for Rheumatism. U.S. Pat. No. 5,403,592, 1990.
- Hills, B. A.; Monds, M. K., Enzymatic identification of the load-bearing boundary lubricant in the joint. Br. J. Rheumatol. 1998, 37, (2), 137-142.
- Oloyede, A., Gudimetla, P., Crawford, R., Hills, B. A., Biomechanical responses of normal and delipidized articular cartilage subjected to varying rates of loading. Connective Tissue Research 2004, 45, (2), 86-93.
- Ethell, M. T.; Hodgson, D. R.; Hills, B. A., The synovial response to exogenous phospholipid (synovial surfactant) injected into the equine radiocarpal joint compared with that to prilocaine, hyaluronan and propylene glycol. New Zealand Veterinary Journal 1999, 47, (4), 128-132.
- Pickard, J. E.; Fisher, J.; Ingham, E.; Egan, J., Investigation into the effects of proteins and lipids on the frictional properties of articular cartilage. Biomaterials 1998, 19, (19), 1807-1812.
- Kawano, T.; Miura, H.; Mawatari, T.; Moro-Oka, T.; Nakanishi, Y.; Higaki, H.; Iwamoto, Y., Mechanical effects of the intraarticular administration of high molecular weight hyaluronic acid plus phospholipid on synovial joint lubrication and prevention of articular cartilage degeneration in experimental osteoarthritis. Arthritis Rheum. 2003, 48, (7), 1923-1929.
- Forsey, R. W.; Fisher, J.; Thompson, J.; Stone, M. H.; Bell, C.; Ingham, E., The effect of hyaluronic acid and phospholipid based lubricants on friction within a human cartilage damage model. Biomaterials 2006, 27, (26), 4581-4590.
- Klein, J., Molecular mechanisms of synovial joint lubrication. J. Proc. Inst. Mech Eng., Part J: J. Eng. Tribology 2006, 220, (8), 691-710.
- Burdick et al., Biological lubricant composition and method of applying lubricant composition. U.S. Pat. No. 6,800,298.
- International patent application publication No. WO2003/000190;
- International patent application publication No. WO2004/047792;
- International patent application publication No. WO2002/078445.
-
- DMPC, which was identified as one preferred component of the liposomal biolubricant composition (when used alone or in combination with DPPC) has saturated, medium-length acyl chains (14 carbons), having a Tm slightly lower than the physiological temperature (Tm=23.2° C. for DMPC-MLV and Tm=˜34° C. for DMPC/DPPC [0.6/1.0 mole/mole] used), thus both PC compositions providing liposomes which are in the LD phase at 37° C. (see Table in Materials and Methods below). When in the LD phase, PC polar headgroup is highly hydrated (˜9.7 water molecules per DMPC or DPPC headgroup, in comparison to <4.3 water molecules per headgroup when below the Tm in the SO phase)53;
- The adiabatic compressibility data presented herein below demonstrate the differences between PC in the solid-ordered (SO) phase (low K values) and the LD phase (higher K values) and the superiority of the LD phase. Partial adiabatic lipid bilayer compressibility (K), which correlates well with the thermotropic behavior54 and was found to reflect the level of hydration, physical state and the volume of cavities (free volume) in the lipid bilayer45. Bound water molecules, which interact with the PC headgroup, are suggested to affect the total volume of cavities in the bilayer, thus affecting intermolecular interactions, as well as the adiabatic compressibility. Specifically, both DOPC and DMPC are in the LD phase (above their Tm, Table 1 below) at (24° C. as well as at 37° C. However, the lubrication ability of DMPC liposomes was substantially superior to that of DOPC. Without being bound by theory, it is believed that the difference in behavior between DMPC and DOPC resides in the fact that under physiological conditions, i.e. at a temperature of between 36° C. and 43° C. DMPC is only slightly above the Tm. Moreover, the temperature in synovial joints of the hand can be as low as ˜28 C. Under such conditions DMPC is also slightly above the Tm. In addition, DMPC is the PC with the shortest acyl chains capable of forming stable liposomes, thus composing the mechanically “softest” bilayer of all other single-component PC bilayers exemplified herein44.
- The lubrication ability of MLV composed of DMPC/DPPC (0.6:1.0 mole/mole) mixture having good miscibility and nearly ideal mixing properties, and a combined SO-to-LD phase transition temperature of ˜34° C. The DMPC/DPPC-MLV showed high lubricating efficacy at 37° C. (static and dynamic friction coefficients of 0.017 and 0.0083, respectively) but not at 24° C. (0.042 and 0.021, respectively), compared with DPPC-MLV alone (Tm of 41.4° C.) which were inferior at 37° C. (0.029 and 0.022, for the static and dynamic friction coefficients, respectively);
- The more efficient lubrication achieved by using liposomes with Tm only slightly below physiological temperature.
- The protection of cartilage from wear using MLV composed of DMPC/DPPC, where a mixture of DMPC/DPPC added to ISF, substantially reduced the wear in comparison to ISF alone and ISF with an addition of HA; this protection effect being significantly higher than the other exemplified membrane, in particular that composed of DPPC alone.
- The “softness” and hydration level of DMPC-MLV and the impact of changes in these features on cartilage lubrication. The first modification in formulation included introduction of ˜33 mole % cholesterol into liposome membranes. As shown below, this resulted in a physical transition from the LD phase to the liquid-ordered (LO) phase34. Such a change is known to “dry” the lipid bilayer56, and is also reflected in a reduction in the adiabatic compressibility and therefore in bilayer softness. Therefore, lubricating cartilage with DMPC/cholesterol-MLV was substantially inferior to lubricating of cartilage with DMPC-MLV (
FIG. 1 ). In another modification 5 mole % of the lipopolymer mPEG-DSPE into the lipid bilayer of DMPC-MLV was introduced. The PEG moieties, extending 4-10 nm from the liposome surface (depending on the polymer chain state, being either in a mushroom or brush configuration39), and are highly flexible and highly hydrated (3 to 4 water molecules per ethylene oxide group)45. However, addition of mPEG-DSPE to DMPC liposomes did not improve lubrication (FIG. 1 ), which seemed to be contradictory to the role of hydration in lubrication. This may be explained by the fact that the PEG moiety although highly polar is nonionic and therefore its hydration differs from that of the hydration of ionic the PC headgroup45. It must be noted, that these grafted PEG moieties may still be beneficial in vivo as they can protect the liposomes from interacting with macromolecules of interstitial fluid34, similarly to the cartilage-protecting behavior of HA22; - Friction coefficients obtained by different media (saline, ISF, and low ionic strength HB) demonstrated that HB was superior to saline and to ISF (
FIGS. 1 , 2, 5-7). Furthermore, the total PL concentration of cartilage specimens lubricated with HB was nearly twice that of cartilage lubricated with ISF and substantially higher than that of cartilage lubricated with saline (FIG. 7 ). Suggesting that HB may better retain naturally-occurring cartilage SAPLs, thereby improving lubrication. The superiority of HB over saline (FIG. 1 ) can also be explained by its lower ionic strength, which induces a less compact PL packing in the lipid bilayer, thus enabling rapid bilayer recovery after frictional events34, 57. This further supports the importance of bilayer softness as a major contributor to effective lubrication. From the above, it became apparent that HB is an effective and supportive medium for liposomes as lubricants; - Large multilamellar DMPC-MLV were found to be superior to small unilamellar liposomes (<100 nm). Without being limited by theory as it is not required for the establishment of the invention, it is believed that this superiority stems from the way the former are retained near the cartilage surface, as demonstrated by the PC distribution along cartilage depth (
FIG. 8 ), due to the large size of MLV (0.8-3.5 μm in diameter). Maroudas et al. reported the presence of 100-nm gaps between collagen fibers in cartilage50. Stockwell and Barnett51 and Barnett and Palfrey52 state that these fibers act as barriers against penetration of large particles into the cartilage, reporting that small silver proteinate particles penetrated deeper than large particles into cartilage. The results presented herein show that smaller DMPC-SUV penetrated deeply into cartilage, while DMPC-MLV remained near the surface (FIG. 8 ). This is in agreement with the similarity of friction levels obtained from cartilage lubricated with DMPC-SUV in HB and of cartilage lubricated with HB alone (FIG. 1 ), as DMPC-SUV penetrate deeply into the cartilage the effect of lubrication is primarily of the media (i.e. HB). - SEM morphological studies, in which naturally-occurring globular structures, in the size range of DMPC-MLV, seemed to be present on the surface of healthy non-lubricated cartilage prior to conducting friction tests (
FIG. 9A ), and absent after friction tests of healthy cartilage lubricated with saline or ISF (FIGS. 9C and 9D , respectively). Cartilage specimens lubricated with DMPC-MLV seemed to have globular lipidic structures on their surface, after conducting friction tests (FIG. 9F ).
TABLE 1 |
phase transition temperatures |
Phase | |||
transition | |||
Phase at | temperature | ||
Lipid | Chemical name (source) | 37° C. | (Tm), ° C. |
HSPC | hydrogenated soybean | SO | 52.5 |
phosphatidylcholine | |||
(Lipoid, Ludwigshafen, Germany) | |||
| 1,2-dipalmitoyl-sn-glycero-3- | SO | 41.4 |
phosphocholine | |||
(Avanti, Alabaster, AL, USA) | |||
| 1,2-dimyristoyl-sn-glycero-3- | LD | 23.2 |
phosphocholine | |||
(Lipoid or Avanti) | |||
| 1,2-dioleoyl-sn-glycero-3- | LD | −21 |
phosphocholine) (Lipoid or Avanti) | |||
Mixture of | LD | 34 | |
DMPC/ | |||
DPPC | |||
(0.6/0.1) | |||
Water: Water was purified using a WaterPro PS HPLC/Ultrafilter Hybrid system (Labconco, Kansas City, Mo.), providing pyrogen-free water with low levels of total carbons and inorganic ions (18.2 MΩ).
Reagents: All other reagents used are of analytical grade or better.
Liposomes: Multilamellar liposomes (MLV) were prepared by dissolving the desired lipids in tert-butanol, followed by lyophilization to form a dry “cake”. This was hydrated in low ionic strength (5 mM) histidine buffer (HB) pH 6.7, at a temperature at least 5° C. above the Tm 34. When desired, MLV were downsized to form small unilamellar vesicles (<100 nm, SUV) by stepwise extrusion through polycarbonate membranes (GE-Osmonics, Minnetonka, Minn.), starting with a 400-nm and ending with a 50-nm-pore-size membrane, using a 10-mL extrusion system (Northern Lipids, Vancouver, Canada) heated at least 5° C. above the Tm 37.
-
- (i) phospholipid (PL) concentration, using the modified Bartlett assay37, 38;
- (ii) size distribution, for liposomes under 1 μm by dynamic light scattering using an ALV-NIBS High Performance Particle Sizer (Langen, Germany) at a scattering angle of 173°; and for liposomes above 400 nm by light diffraction using a Beckman Coulter LS Particle Size Analyzer 13-320 (Fullerton, Calif.), equipped with polarization intensity differential scattering (PIDS) to provide a dynamic detection range from 40 nm to 2000 μm;
- (iii) partial specific adiabatic compressibility, by calculation from the density of the liposome dispersion (using a DMA 5000 density meter, Anton Paar, Graz, Austria) and the velocity of an 5 MHz ultrasonic wave traveling through it (using a UCC-12 ultrasonic velocimeter, NDT Instruments, Jerusalem, Israel), as described by Garbuzenko et al.39; and
- (iv) structure, using scanning electron microscopy (SEM).
Cartilage: Articular cartilage from healthy or osteoarthritis humans (aged 65 to 86 years) was obtained from femoral head fracture operations or total hip replacements. Tissue was classified as normal or pathological according to the visual diagnosis. For wear rate determination, only femoral heads with normal tissue were selected. Specimens were frozen at −20° C. until sample preparation in order to keep the mechanical properties close to those of live tissue. Pairs of cylindrical plugs, having 4 mm and 8 mm in diameter, each pair from the same region of the joint, were prepared. These cylindrical plugs, consisting of about 2 mm thick cartilage on top of about 8 mm long bone, were removed from the femoral head using a cork borer. The plugs were glued to holders through the bone part, using cyanoacrylate-based adhesive glue, leaving the cartilage projecting out of the holders. Thereafter, these plugs were refrozen at −20° C. until tested. Cartilage with completely intact and smooth surface was used. Friction and wear testing: Liposomes covering a wide range of sizes and concentrations, dispersed in HB, were screened as potential lubricants to reduce friction and wear between two discs of human cartilage at 24° C. and 37° C. Friction measurements were carried out with a cartilage-on-cartilage setup (Merkher, Y.; Sivan, S.; Etsion, I.; Maroudas, A.; Halperin, G.; Yosef, A., A rational human joint friction test using a human cartilage-on-cartilage arrangement. Tribol. Lett. 2006, 22, 29-36, the content of which is incorporated herein by reference in its entirety), using two discs of cartilage immersed in a liposomal dispersion in HB, or as controls, in HB alone, or in physiological saline (0.9% w/v; pH 5.0; Teva Medical, Israel), or in inflamed synovial fluid (ISF) obtained from osteoarthritis patients. These discs were subjected to relative sliding over a wide range of loads (1 to 30 N), equivalent to physiological pressures in joints (0.08 to 2.4 MPa). Various sliding velocities (0.5 to 2 mm/s) and dwell times (5 to 300 s) were used to simulate, together with various loads, a range of physiological movements.
For the a qualitative evaluation of wear, the effect of friction tests on the concentration of total PL in cartilage, and on the structure of the cartilage surface was determined.
PL extraction and quantification: Total PL were extracted from cartilage specimens before and after lubrication tests, using the Bligh and Dyer extraction procedure41, 42. For this, cartilage specimens were incubated in a chloroform-methanol solution (1:1 v/v) for 1 h at 37° C. Water was added to a final chloroform-water-methanol ratio of 1:1:1, the solution was Vortexed for 1 min and then centrifuged, using a desk centrifuge, to form two phases. The chloroform-rich lower phase, containing the PL, was collected, dried under vacuum (Concentrator 5301, Eppendorf), and the residual (containing lipids) was re-dissolved in a small volume of chloroform-methanol solution (2:1 v/v) and then loaded onto low-phosphorus silica gel TLC glass plates (Uniplate—Silica Gel G, Analtech, Newark, Del.). A chloroform-methanol-water (65:25:4 v/v/v) solvent system was used for TLC41. Commercial markers of sphingomyelin, PC and PE were also loaded on the plates for spot identification. Lipid spots were detected after spraying the dried TLC plates with a UV-detectable primulin (Sigma) solution (1 mL of 0.1% w/v primulin in water, added to 100 mL acetone-water, 4:1 v/v). Each PL spot was scraped from the TLC plate, and its PL content was quantified by the modified Bartlett procedure.37, 38
Results
Liposome carrier: it was found that the lubrication efficiency of HB is better than that of saline, or of ISF (
Effect of Liposome Size and Lamellarity on their Penetration into Cartilage: PC concentration, as a function of cartilage depth (0-800 μm, in 20-50-μm increments), was measured after friction tests for specimens lubricated with DMPC-MLV and DMPC-SUV, both dispersed in HB, and for specimens lubricated with HB alone (control). Among these specimens, cartilage lubricated with DMPC-MLV had the highest PC concentration near the cartilage surface (
Cartilage morphology: SEM was used to study cartilage surface morphology and wear28. In
TABLE 2 |
Group composition, treatments and Identification of animals |
Animal No. | Cage No. | Tail No. | Left | Right Knee | ||||
1, 10, 19, 28, | 1, 4, 7, 10, | 1 | HB | DMPC | ||||
37 | 13 | |||||||
2, 11, 20, 29, | 2 | HB | DMPC | |||||
38 | ||||||||
3, 12, 21, 30, | DMPC | |||||||
39 | ||||||||
4, 13, 22, 31, | 2, 5, 8, 11, | 1 | Physiological | DMPC + | ||||
40 | 14 | | DPPC | |||||
5, 14, 23, 32, | 2 | Physiological | DMPC + | |||||
41 | | DPPC | ||||||
6, 15, 24, 33, | HB | DMPC + | ||||||
42 | | |||||||
7, 16, 25, 34, | 3, 6, 9, 12 | 1 | DMPC | DMPC | ||||
43 | 15 | |||||||
8, 17, 26, 35, | 15 | 2 | DMPC + DPPC | DMPC | ||||
44 | ||||||||
9, 18, 27, 36, | 4 | DMPC + DPPC | DMPC | |||||
45 | DMPC + DPPC | DMPC | ||||||
46 | ||||||||
*Physiological saline: 0.9% NaCl |
Test compositions: Liposomal DMPC and DMPC+DPPC were prepared as described above (the liposomes being dispersed in HB) and stored in 4° C. till one hour before the application. The last hour before the application they were left in room temperature. The concentrations of the liposomes were for DPPC 100.6 mM and for DMPC+DPPC 92.0 mM.
TABLE 3 |
Table of weights and treatments |
Cage | Tail | Initial | |||||||
Rat No. | No. | No. | Left knee | Right Knee | Weight | 2nd weight | Final Weight | 2nd treatment | Eutanization |
1 | 1 | 1 | HB | DMPC | 298 | 303 | No | day (19/12) | |
2 | 1 | 2 | HB | DMPC | 310 | 316 | |||
3 | 1 | 3 | Physiological saline | DMPC | 300 | 295 | |||
4 | 2 | 1 | Physiological saline | DMPC + DPPC | 310 | 318 | |||
5 | 2 | 2 | Physiological saline | DMPC + DPPC | 298 | 285 | |||
6 | 2 | HB | DMPC + DPPC | 290 | 292 | ||||
7 | 3 | 1 | DMPC + DPPC | DMPC | 285 | 277 | |||
8 | 3 | 2 | DMPC + DPPC | DMPC | 290 | 295 | |||
9 | 3 | DMPC + DPPC | DMPC | 290 | 286 | ||||
10 | 4 | 1 | HB | DMPC | 302 | 307 | No | 4 days (22/12) | |
11 | 4 | 2 | HB | DMPC | 303 | 324 | |||
12 | 4 | Physiological saline | DMPC | 298 | 317 | ||||
13 | 5 | 1 | Physiological saline | DMPC + DPPC | 315 | 330 | |||
14 | 5 | 2 | Physiological saline | DMPC + DPPC | 314 | 330 | |||
15 | 5 | HB | DMPC + DPPC | 282 | 292 | ||||
16 | 6 | 1 | DMPC + DPPC | DMPC | 280 | 280 | |||
17 | 6 | 2 | DMPC + DPPC | DMPC | 305 | 305 | |||
18 | 6 | DMPC + DPPC | DMPC | 298 | 298 | ||||
19 | 7 | 1 | HB | DMPC | 312 | 312 | No | fortnight (1/1) | |
20 | 7 | 2 | HB | DMPC | 299 | 299 | |||
21 | 7 | Physiological saline | DMPC | 318 | 318 | ||||
22 | 8 | 1 | Physiological saline | DMPC + DPPC | 303 | 303 | |||
23 | 8 | 2 | Physiological saline | DMPC + DPPC | 300 | 300 | |||
24 | 8 | HB | DMPC + DPPC | 300 | 300 | ||||
25 | 9 | 1 | DMPC + DPPC | DMPC | 300 | 300 | |||
26 | 9 | 2 | DMPC + DPPC | DMPC | 321 | 321 | |||
27 | 9 | DMPC + DPPC | DMPC | 300 | 300 | ||||
28 | 10 | 1 | HB | DMPC | 313 | 405 | No | 4 weeks (15/1) | |
29 | 10 | 2 | HB | DMPC | 310 | 409 | |||
30 | 10 | Physiological saline | DMPC | 298 | 318 | ||||
31 | 11 | 1 | Physiological saline | DMPC + DPPC | 289 | 356 | |||
32 | 11 | 2 | Physiological saline | DMPC + DPPC | 298 | 364 | |||
33 | 11 | HB | DMPC + DPPC | 308 | 404 | ||||
34 | 12 | 1 | DMPC + DPPC | DMPC | 290 | 355 | |||
35 | 12 | 2 | DMPC + DPPC | DMPC | 312 | 420 | |||
36 | 12 | DMPC + DPPC | DMPC | 315 | 389 | ||||
37 | 13 | 1 | HB | DMPC | 306 | 361 | 400 | fortnight 1/1 | 4 weeks (15/1) |
38 | 13 | 2 | HB | DMPC | 298 | 342 | 374 | ||
39 | 13 | Physiological saline | DMPC | 300 | 338 | 359 | |||
40 | 14 | 1 | Physiological saline | DMPC + DPPC | 301 | 351 | 383 | ||
41 | 14 | 2 | Physiological saline | DMPC + DPPC | 298 | 341 | 355 | ||
42 | 14 | HB | DMPC + DPPC | 300 | 338 | 368 | |||
43 | 15 | 1 | DMPC + DPPC | DMPC | 306 | 348 | 384 | ||
44 | 15 | 2 | DMPC + DPPC | DMPC | 291 | 336 | 337 | ||
45 | 15 | DMPC + DPPC | DMPC | 298 | 338 | 384 | |||
46 | 15 | 4 | DMPC + DPPC | DMPC | 298 | 351 | 352 | ||
Clinical Evaluation: The animal technician held physical observations on the rats throughout the study for monitoring prominent change in their conditions such as drastic weight loss, wounds or deaths. All rats were weighted before the applications and after euthanization.
Necropsy: All rats were subjected to a full detailed necropsy and gross pathological examination at the end of the study period following euthanasia. At necropsy, all rats were thoroughly examined for any abnormality or gross pathological changes in tissues and/or organs. Samples of all knees were sent to slides preparation at Patho-Lab Ltd. Ness Ziona. Tissue fixation was obtained in formaldehyde (Bio-Lab Ltd.).
- 1. Alexander, C. J. Idiopathic osteoarthritis: time to change paradigms? Skeletal Radiol. 33, 321-324 (2004).
- 2. Corti, M. C. & Rigon, C. Epidemiology of osteoarthritis: prevalence, risk factors and functional impact. Aging Clin. Exp. Res. 15, 359-363 (2003).
- 3. Bullough, P. G. & Vigorita, V. J. Orthopaedic Pathology, Edn. 3rd. (Mosby-Wolfe, Baltimore; 1997).
- 4. Koopman, W. J. & Moreland, L. W. Arthritis and Allied Conditions: A Textbook of Rheumatology, Vol. 1-2, Edn. 15th. (Lippincott Williams & Wilkins, Philadelphia; 2005).
- 5. Sokoloff, L. The biology of degenerative joint disease. Acta Rhumatol Belg. 1, 155-156 (1977).
- 6. Conaghan, P. G., Vanharanta, H. & Dieppe, P. A. Is progressive osteoarthritis an atheromatous vascular disease? Ann. Rheum. Dis. 64, 1539-1541 (2005).
- 7. Neame, R. & Doherty, M. Osteoarthritis update. Clin. Med. 5, 207-210 (2005).
- 8. Imhof, H. et al. Subchondral bone and cartilage disease: a rediscovered functional unit. Invest. Radial. 35, 581-588 (2000).
- 9. Lajeunesse, D. & Reboul, P. Subchondral bone in osteoarthritis: a biologic link with articular cartilage leading to abnormal remodeling. Curr. Opin. Rheumatol. 15, 628-633 (2003).
- 10. Radin, E. L. Who gets osteoarthritis and why? J. Rheumatol. Suppl. 70, 10-15 (2004).
- 11. Grainger, R. & Cicuttini, F. M. Medical management of osteoarthritis of the knee and hip joints. Med. J. Aust. 180, 232-236 (2004).
- 12. Swaim, D. A., Hendren, R. B., Radin, E. L., Sotman, S. L. & Duda, B. A. The lubricating activity of synovial fluid glycoproteins. Arthritis Rheum, 24, 22-30 (1981).
- 13. Swarm, D. A. & Mintz, G. The isolation and properties of a second glycoprotein (LGP-II) from the articular lubricating fraction from bovine synovial fluid. Biochem. J. 179, 465-471 (1979).
- 14. Swami, D. A., Slayter, H. S. & Silver, F. H. The molecular structure of lubricating glycoprotein-1, the boundary lubricant for articular cartilage. J. Biol. Chem. 256, 5921-5925 (1981).
- 15. Nitzan, D. W., Kreiner, B. & Zeltser, R. TMJ lubrication system: its effect on the joint function, dysfunction, and treatment approach. Compend. Contin. Educ. Dent. 25, 437-444 (2004).
- 16. Yui, N., Okano, T. & Sakurai, Y. Inflammation responsive degradation of crosslinked hyaluronic acid gels. J. Control. Release 22, 105-116 (1992).
- 17. Hills, B. A. & Butler, B. D. Surfactants identified in synovial fluid and their ability to act as boundary lubricants. Ann. Rheum. Dis. 43, 641-648 (1984),
- 18. Sarma, A. V., Powell, G. L. & LaBerge, M. Phospholipid composition of articular cartilage boundary lubricant. J. Orthop. Res. 19, 671-676 (2001).
- 19. Schwarz, I. M. & Hills, B. A. Surface-active phospholipid as the lubricating component of lubricin. Br. J. Rheumatol. 37, 21-26 (1998).
- 20. Hills, B. A. & Monds, M. K. Enzymatic identification of the load-bearing boundary lubricant in the joint. Br. J. Rheumatol. 37, 137-142 (1998).
- 21. Ogston, A. G. & Stanier, J. E. Physiological function of hyaluronic acid in synovial fluid; viscous, elastic, and lubricant properties. J. Physiol. (Cambridge) 119, 244-252 (1953).
- 22. Benz, M., Chen, N. & Israelachvili, J. Lubrication and wear properties of grafted polyelectrolytes, hyaluronan and hylan, measured in the surface forces apparatus. J. Biomed. Mater. Res. A. 71, 6-15 (2004).
- 23. Rhee, D. K. et al, The secreted glycoprotein lubricin protects cartilage surfaces and inhibits synovial cell overgrowth. J. Clin. Invest. 115, 622-631 (2005).
- 24. Swann, D. A., Bloch, K. J., Swindell, D. & Shore, E. The lubricating activity of human synovial fluids. Arthritis Rheum. 27, 552-556 (1984).
- 25. Pickard, J. E., Fisher, J., Ingham, E. & Egan, J. Investigation into the effects of proteins and lipids on the frictional properties of articular cartilage. Biomaterials 19, 1807-1812 (1998).
- 26. Vecchio, P., Thomas, R. & Hills, B. A. Surfactant treatment for osteoarthritis. Rheumatology (Oxford) 38, 1020-1021 (1999).
- 27. Gudimelta, O. A., Crawford, R. & Hills, B. A. Consilidation responses of delipidized cartilage. Clin. Biomech. 19, 534-542 (2004).
- 28. Watanabe, M. et al. Ultrastructural study of upper surface layer in rat articular cartilage by “in vivo cryotechnique” combined with various treatments. Med. Elect. Microsc. 33, 16-24 (2000).
- 29. Kawano, T. et al. Mechanical effects of the intraarticular administration of high molecular weight hyaluronic acid plus phospholipid on synovial joint lubrication and prevention of articular cartilage degeneration in experimental osteoarthritis. Arthritis Rheum. 48, 1923-1929 (2003).
- 30. Forsey, R. W. et al. The effect of hyaluronic acid and phospholipid based lubricants on friction within a human cartilage damage model. Biomaterials 27, 4581-4590 (2006).
- 31. Klein, J. Molecular mechanisms of synovial joint lubrication. J. Proc. Inst. Mech Eng., Part J: J. Eng. Tribology 220, 691-710 (2006).
- 32. Briscoe, W. H. et al. Boundary lubrication under water. Nature 444, 191-194 (2006).
- 33. Raviv, U. et al. Lubrication by charged polymers. Nature 425, 163-165 (2003).
- 34. Barenholz, Y. & Cevc, G. Structure and properties of membranes in Physical Chemistry of Biological Surfaces. (Marcel Dekker, New York; 2000).
- 35. Israelachvili, J., Intermolecular and surface Forces, 2nd edition, Academic Pres, London (1992)
- 36. Marsh, D. CRC Handbook of Lipid Bilayers. (CRC Press, Boca Raton, Fla.; 1990).
- 37. Barenholz, Y. & Amselem, S. Quality control assays in the development and clinical use of liposome-based formulations in Liposome Technology, Edn. 2nd, (CRC, Boca Raton, Fla.; 1993).
- 38. Shmeeda, H., Even-Chen, S. & Barenholz, Y. Enzymatic assays for quality control and pharmacokinetics of liposome formulations: Comparison with nonenzymatic conventional methodologies. Methods Enzymol. 367, 272-292 (2003).
- 39. Garbuzenko, O., Barenholz, Y. & Priev, A. Effect of grafted PEG on liposome size and on compressibility and packing of lipid bilayer. Chem. Phys. Lipids 135, 117-129 (2005).
- 40. Merkher, Y. et al. A rational human joint friction test using a human cartilage-on-cartilage arrangement. Tribol. Lett. 22, 29-36 (2006).
- 41. Barenholz, Y. Relevancy of drug loading to liposomal formulation therapeutic efficacy. J. Liposome Res. 13, 1 (1993).
- 42. Bligh, E. G. & Dyer, W. J. A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol, 37, 911-917 (1959).
- 43. Biltonen, R. L. & Lichtenberg, D. The use of differential scanning calorimetry as a tool to characterize liposome preparations. Chem. Phys. Lipids 64, 129-142 (1993).
- 44. Mouritsen, O. G. Life As a Matter of Fat. The Emerging Science of Lipidomics. (Springer-Verlag, Berlin; 2005).
- 45. Tirosh, O., Barenholz, Y., Katzhendler, J. & Priev, A. Hydration of polyethylene glycol-grafted liposomes. Biophys. J. 74, 1371-1379 (1998).
- 46. Hills, B. A. Boundary lubrication in vivo. J. Eng. Med. 214, 83-94 (2000).
- 47. Mabrey, S. & Sturtevant, J. M. Investigation of phase transitions of lipids and lipid mixtures by high sensitivity differential scanning calorimetry. PNAS 73, 3862-3866 (1976).
- 48. Yoshida, M., Zea-Aragon, Z., Ohtsuki, K., Ohnishi, M. & Ohno, S. Ultrastructural study of upper surface layer in rat mandibular condylar cartilage by quick-freezing method. Histol. Histopathol. 19, 1033-1041 (2004).
- 49. Klein, J. Mechanism of friction across molecularly confined films of simple liquids. Tribology Series 36, 59-64 (1999).
- 50. Maroudas, A. Distribution and diffusion of solutes in articular cartilage. Biophys. J. 10, 365-379 (1970).
- 51. Stockwell, R. A. & Bartlett, C. H. Changes in permeability of articular cartilage with age. Nature 201, 835-836 (1964).
- 52. Barnett, C. H. & Palfrey, A. J. Absorption into the rabbit articular cartilage. J. Anat. 99, 365-375 (1965).
- 53. Faure, C., Bonakdar, L. & Dufourc, E. J. Determination of DMPC hydration in the L(alpha) and L(beta′) phases by 2H solid state NMR of D2O. FEBS Lett. 405, 263-266 (1997).
- 54. Schrader, W. et al. Compressibility of lipid mixtures studied by calorimetry and ultrasonic velocity measurements. J. Phys. Chem. B 106, 6581-6586 (2002).
- 55. Schwarz, U.S., Komura, S. & Safran, S. A. Deformation and tribology of multi-walled hollow nanoparticles. Europhys. Lett. 50, 762-768 (2000).
- 56. Parasassi, T., Di Stefano, M., Loiero, M., Ravagnan, G. & Gratton, E. Cholesterol modifies water concentration and dynamics in phospholipid bilayers: a fluorescence study using Laurdan probe. Biophys J. 66, 763-768 (1994).
- 57. Oncins, G., Garcia-Manyes, S. & Sanz, F. Study of frictional properties of a phospholipid bilayer in a liquid environment with lateral force microscopy as a function of NaCl concentration. Langmuir 21, 7373-7349 (2005).
- 58. Ballantine G. C., Stachowiak G. W., The effects of lipid depletion on osteoarthritic wear, Wear 253, 385-393 (2002).
- 59 Jones C. F., Stoffel K., Ozturk H. E., Stachowiak G. W., The effect of surface active phospholipids on the lubrication of osteoarthritic sheep knee joints: wear, Tribal. Lett. 16(4), 291-296 (2004).
- 60 Hills B. A., Monds M. K., Deficiency of lubricating surfactant lining the articular surfaces of replaced hips and knees, Br. J. Rheumatol. 37, 143-147 (1998).
- 61 Freeman M. A. R. (Ed.), Adult Articular Cartilage, 2nd ed., Chapter 3 Pitman Medical, London (1979).
- 62 Farndale R. W., Buttle D J., Barrett A. J., Improved quantification and discrimination of sulfated glycosaminoglycans by use of dimethylmethylene, Biochim Biophys Acta 883(2), 173-177 (1986).
- 63. International patent application publication No. WO2003/000190;
- 64. International patent application publication No. WO2004/047792;
- 65. International patent application publication No. WO2002/078445
Claims (17)
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US12/411,855 US8895054B2 (en) | 2006-09-28 | 2009-03-26 | Methods for joint lubrication and cartilage wear prevention making use of glycerophospholipids |
US14/520,542 US20150037404A1 (en) | 2006-09-28 | 2014-10-22 | Methods for joint lubrication and cartilage wear prevention making use of glycerophospholipids |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US84765106P | 2006-09-28 | 2006-09-28 | |
PCT/IL2007/001215 WO2008038292A2 (en) | 2006-09-28 | 2007-10-07 | Use of glycerophospholipids for joint lubrication |
US12/411,855 US8895054B2 (en) | 2006-09-28 | 2009-03-26 | Methods for joint lubrication and cartilage wear prevention making use of glycerophospholipids |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/IL2007/001215 Continuation-In-Part WO2008038292A2 (en) | 2006-09-28 | 2007-10-07 | Use of glycerophospholipids for joint lubrication |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US14/520,542 Continuation US20150037404A1 (en) | 2006-09-28 | 2014-10-22 | Methods for joint lubrication and cartilage wear prevention making use of glycerophospholipids |
Publications (2)
Publication Number | Publication Date |
---|---|
US20100098749A1 US20100098749A1 (en) | 2010-04-22 |
US8895054B2 true US8895054B2 (en) | 2014-11-25 |
Family
ID=39144533
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US12/411,855 Active 2029-03-25 US8895054B2 (en) | 2006-09-28 | 2009-03-26 | Methods for joint lubrication and cartilage wear prevention making use of glycerophospholipids |
US14/520,542 Abandoned US20150037404A1 (en) | 2006-09-28 | 2014-10-22 | Methods for joint lubrication and cartilage wear prevention making use of glycerophospholipids |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US14/520,542 Abandoned US20150037404A1 (en) | 2006-09-28 | 2014-10-22 | Methods for joint lubrication and cartilage wear prevention making use of glycerophospholipids |
Country Status (8)
Country | Link |
---|---|
US (2) | US8895054B2 (en) |
EP (1) | EP2079442B1 (en) |
JP (1) | JP5660561B2 (en) |
CN (1) | CN101541308B (en) |
CA (1) | CA2664944C (en) |
ES (1) | ES2599630T3 (en) |
IL (1) | IL197827A (en) |
WO (1) | WO2008038292A2 (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20130142863A1 (en) * | 2010-06-17 | 2013-06-06 | Yissum Research Development Company Of The Hebrew University Of Jerusalem, Ltd. | Phosphatidylcholine lipid liposomes as boundry lubricants in aqueous media |
US20200170948A1 (en) * | 2017-08-22 | 2020-06-04 | Moebius Medical Ltd. | Liposomal formulation for joint lubrication |
Families Citing this family (22)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2004096140A2 (en) | 2003-04-25 | 2004-11-11 | The Penn State Research Foundation | Method and system for systemic delivery of growth arresting, lipid-derived bioactive compounds |
EP2079442B1 (en) * | 2006-09-28 | 2016-07-27 | Hadasit Medical Research Services & Development Limited | Use of glycerophospholipids for joint lubrication |
JP2010189315A (en) * | 2009-02-18 | 2010-09-02 | Unitika Ltd | Healing accelerator for articular cartilage lesion and food and drink containing the same |
MY170121A (en) * | 2009-06-03 | 2019-07-05 | Ulrich Vierl | Formulations for the treatment of deep tissue pain |
BR112012003834A2 (en) | 2009-08-21 | 2017-08-08 | Targeted Delivery Tech Limited | methods for treating disorders and for treating fatty acid deficiency, hypertriglyceridemia or hypercholesterolemia and package |
US8716204B2 (en) | 2010-07-27 | 2014-05-06 | Zimmer, Inc. | Synthetic synovial fluid compositions and methods for making the same |
RU2480174C1 (en) * | 2011-11-22 | 2013-04-27 | Сергей Талустанович Сохов | Method of treating diseases of temporomandibular joint, accompanied by anterior displacement and deformation of articular disk |
GB201205642D0 (en) | 2012-03-29 | 2012-05-16 | Sequessome Technology Holdings Ltd | Vesicular formulations |
GB201206486D0 (en) * | 2012-04-12 | 2012-05-30 | Sequessome Technology Holdings Ltd | Vesicular formulations and uses thereof |
US10653786B2 (en) | 2012-04-25 | 2020-05-19 | Trustees Of Tufts College | Silk microspheres and methods for surface lubrication |
WO2015001564A1 (en) | 2013-07-04 | 2015-01-08 | Yeda Research And Development Co. Ltd. | Low friction hydrogels and hydrogel-containing composite materials |
US9982027B2 (en) | 2013-10-22 | 2018-05-29 | Lubris Llc | Control of rheological properties of mixed hyaluronate/lubricin solutions |
ES2897733T3 (en) | 2014-06-15 | 2022-03-02 | Yeda Res & Dev | Treatment of contact lens surfaces and treatment of ocular discomfort by means of water-soluble polymers and lipids/liposomes |
US10646574B2 (en) | 2014-07-21 | 2020-05-12 | Board Of Regents, The University Of Texas System | Formulations of intraarticular pharmaceutical agents and methods for preparing and using the same |
IL234929B (en) | 2014-10-01 | 2021-01-31 | Yeda Res & Dev | Liposomes-containing antifouling compositions and uses thereof |
EP3954361A1 (en) | 2015-06-30 | 2022-02-16 | Sequessome Technology Holdings Limited | Multiphasic compositions |
RU2675343C1 (en) * | 2018-03-22 | 2018-12-18 | Валерий Валентинович Бекреев | Method of minimally invasive surgical treatment of internal discrepancy of temporomandibular joint, followed by slipped articular disk, with arthrocentesis |
CN110314137B (en) * | 2018-03-30 | 2022-04-05 | 北京泰德制药股份有限公司 | Freeze-dried preparation containing lipid vesicles |
CN108743524B (en) * | 2018-06-12 | 2020-10-30 | 清华大学 | Application of liposome in lubricating liquid, lubricating liquid and preparation method thereof |
TWI782181B (en) * | 2019-02-11 | 2022-11-01 | 以色列商莫比斯醫療有限公司 | Liposomal formulation for joint lubrication |
US11607386B2 (en) * | 2019-04-03 | 2023-03-21 | New York University | Liposomes encapsulating adenosine |
WO2023048097A1 (en) * | 2021-09-21 | 2023-03-30 | 株式会社アイシン | Lipid membrane endoplasmic reticulum, fabrication device, and fabrication method for same |
Citations (16)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5304380A (en) * | 1989-11-09 | 1994-04-19 | Japan Tobacco Inc. | Glucosamine derivative and liposome containing the same as membrane constituent |
US5403592A (en) | 1987-08-25 | 1995-04-04 | Macnaught Pty Limited | Lubricant composition for rheumatism |
US6133249A (en) | 1995-12-19 | 2000-10-17 | Macnaught Medical Pty Limited | Phospholipid and propylene glycol based lubricant |
WO2002078445A1 (en) | 2001-03-29 | 2002-10-10 | The Scripps Research Institute | Formulations comprising entrapped active ingredients and uses thereof |
US6538032B1 (en) * | 2001-09-07 | 2003-03-25 | Charmzone Co., Ltd. | Phytosphingosine derivatives with antitumor activity |
US20030139344A1 (en) * | 2001-11-19 | 2003-07-24 | Mien-Chie Hung | Antitumor activity of Bok |
US6800298B1 (en) | 2000-05-11 | 2004-10-05 | Clemson University | Biological lubricant composition and method of applying lubricant composition |
US20050069576A1 (en) * | 2003-09-25 | 2005-03-31 | Robert Mills | Mahonia aquifolium extract, extraction process and pharmaceutical composition containing the same |
US20050123593A1 (en) * | 2001-06-25 | 2005-06-09 | Jonathan Thompson | Liposomal encapsulation of glycosaminoglycans for the treatment of arthritic joints |
US20060029655A1 (en) | 2001-06-25 | 2006-02-09 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | Method for preparation of vesicles loaded with biological material and different uses thereof |
US20060147511A1 (en) | 2002-11-24 | 2006-07-06 | Steffen Panzner | Liposomal glucocorticoids |
US20100098749A1 (en) * | 2006-09-28 | 2010-04-22 | Hadasit Medical Research Services & Development Limited | Methods for joint lubrication and cartilage wear prevention making use of glycerophospholipids |
US7749485B2 (en) * | 2004-06-03 | 2010-07-06 | Bracco Research S.A. | Liposomal assembly for therapeutic and/or diagnostic use |
US20110171288A1 (en) * | 2008-02-20 | 2011-07-14 | Fatemeh Mohammadi | Topical compositions and methods for whitening skin |
US20120213844A1 (en) * | 2006-12-21 | 2012-08-23 | Ken Shi Kun Huang | Liposome composition for targeting egfr receptor |
US20140079774A1 (en) * | 2011-04-28 | 2014-03-20 | Stc.Unm | Porous nanoparticle-supported lipid bilayers (protocells) for targeted delivery and methods of using same |
-
2007
- 2007-10-07 EP EP07827189.7A patent/EP2079442B1/en active Active
- 2007-10-07 CN CN2007800440724A patent/CN101541308B/en active Active
- 2007-10-07 ES ES07827189.7T patent/ES2599630T3/en active Active
- 2007-10-07 CA CA2664944A patent/CA2664944C/en active Active
- 2007-10-07 JP JP2009529856A patent/JP5660561B2/en active Active
- 2007-10-07 WO PCT/IL2007/001215 patent/WO2008038292A2/en active Application Filing
-
2009
- 2009-03-26 IL IL197827A patent/IL197827A/en active IP Right Grant
- 2009-03-26 US US12/411,855 patent/US8895054B2/en active Active
-
2014
- 2014-10-22 US US14/520,542 patent/US20150037404A1/en not_active Abandoned
Patent Citations (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5403592A (en) | 1987-08-25 | 1995-04-04 | Macnaught Pty Limited | Lubricant composition for rheumatism |
US5304380A (en) * | 1989-11-09 | 1994-04-19 | Japan Tobacco Inc. | Glucosamine derivative and liposome containing the same as membrane constituent |
US6133249A (en) | 1995-12-19 | 2000-10-17 | Macnaught Medical Pty Limited | Phospholipid and propylene glycol based lubricant |
US6800298B1 (en) | 2000-05-11 | 2004-10-05 | Clemson University | Biological lubricant composition and method of applying lubricant composition |
US20050164981A1 (en) * | 2000-05-11 | 2005-07-28 | Burdick Julie-Anne M. | Biological lubricant composition and method of applying lubricant composition |
WO2002078445A1 (en) | 2001-03-29 | 2002-10-10 | The Scripps Research Institute | Formulations comprising entrapped active ingredients and uses thereof |
US20050123593A1 (en) * | 2001-06-25 | 2005-06-09 | Jonathan Thompson | Liposomal encapsulation of glycosaminoglycans for the treatment of arthritic joints |
US20060029655A1 (en) | 2001-06-25 | 2006-02-09 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | Method for preparation of vesicles loaded with biological material and different uses thereof |
US6538032B1 (en) * | 2001-09-07 | 2003-03-25 | Charmzone Co., Ltd. | Phytosphingosine derivatives with antitumor activity |
US20030139344A1 (en) * | 2001-11-19 | 2003-07-24 | Mien-Chie Hung | Antitumor activity of Bok |
US20060147511A1 (en) | 2002-11-24 | 2006-07-06 | Steffen Panzner | Liposomal glucocorticoids |
US20050069576A1 (en) * | 2003-09-25 | 2005-03-31 | Robert Mills | Mahonia aquifolium extract, extraction process and pharmaceutical composition containing the same |
US7749485B2 (en) * | 2004-06-03 | 2010-07-06 | Bracco Research S.A. | Liposomal assembly for therapeutic and/or diagnostic use |
US20100098749A1 (en) * | 2006-09-28 | 2010-04-22 | Hadasit Medical Research Services & Development Limited | Methods for joint lubrication and cartilage wear prevention making use of glycerophospholipids |
US20120213844A1 (en) * | 2006-12-21 | 2012-08-23 | Ken Shi Kun Huang | Liposome composition for targeting egfr receptor |
US20110171288A1 (en) * | 2008-02-20 | 2011-07-14 | Fatemeh Mohammadi | Topical compositions and methods for whitening skin |
US20140079774A1 (en) * | 2011-04-28 | 2014-03-20 | Stc.Unm | Porous nanoparticle-supported lipid bilayers (protocells) for targeted delivery and methods of using same |
Non-Patent Citations (58)
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20130142863A1 (en) * | 2010-06-17 | 2013-06-06 | Yissum Research Development Company Of The Hebrew University Of Jerusalem, Ltd. | Phosphatidylcholine lipid liposomes as boundry lubricants in aqueous media |
US11541008B2 (en) * | 2010-06-17 | 2023-01-03 | Yeda Research And Development Co., Ltd. | Phosphatidylcholine lipid liposomes as boundary lubricants in aqueous media |
US20200170948A1 (en) * | 2017-08-22 | 2020-06-04 | Moebius Medical Ltd. | Liposomal formulation for joint lubrication |
US11123293B2 (en) * | 2017-08-22 | 2021-09-21 | Moebius Medical Ltd. | Liposomal formulation for joint lubrication |
EP4223284A1 (en) | 2017-08-22 | 2023-08-09 | Moebius Medical Ltd. | Liposomal formulation for joint lubrication |
Also Published As
Publication number | Publication date |
---|---|
EP2079442A2 (en) | 2009-07-22 |
CN101541308B (en) | 2012-02-01 |
JP2010540406A (en) | 2010-12-24 |
IL197827A0 (en) | 2009-12-24 |
EP2079442B1 (en) | 2016-07-27 |
WO2008038292A2 (en) | 2008-04-03 |
US20150037404A1 (en) | 2015-02-05 |
IL197827A (en) | 2016-02-29 |
US20100098749A1 (en) | 2010-04-22 |
WO2008038292A3 (en) | 2009-02-12 |
ES2599630T3 (en) | 2017-02-02 |
CN101541308A (en) | 2009-09-23 |
CA2664944C (en) | 2016-06-14 |
JP5660561B2 (en) | 2015-01-28 |
CA2664944A1 (en) | 2008-04-03 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US8895054B2 (en) | Methods for joint lubrication and cartilage wear prevention making use of glycerophospholipids | |
Sivan et al. | Liposomes act as effective biolubricants for friction reduction in human synovial joints | |
Verberne et al. | Liposomes as potential biolubricant additives for wear reduction in human synovial joints | |
US11541008B2 (en) | Phosphatidylcholine lipid liposomes as boundary lubricants in aqueous media | |
AU2023203309A1 (en) | Liposomal formulation for joint lubrication | |
TWI782181B (en) | Liposomal formulation for joint lubrication | |
EA042384B1 (en) | LIPOSOMAL COMPOSITION TO PROVIDE A JOINT LUBRICANT | |
BR122022020806B1 (en) | PHARMACEUTICAL COMPOSITION COMPRISING LIPOSOMES, PROCESS FOR PREPARING THE SAME AND USES OF SAID COMPOSITION TO LUBRICATE A JOINT AND TREAT PAIN OR IRRITATION IN A JOINT | |
Schroeder et al. | Surface Active Phospholipids as Cartilage Lubricants |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AS | Assignment |
Owner name: TECHNION RESEARCH AND DEVELOPMENT FOUNDATION LTD., Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:BARENHOLZ, YECHEZKEL;NITZAN, DORIT;ETSION, IZHAK;AND OTHERS;SIGNING DATES FROM 20090527 TO 20090804;REEL/FRAME:024081/0826 Owner name: HADASIT MEDICAL RESEARCH SERVICES & DEVELOPMENT LI Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:BARENHOLZ, YECHEZKEL;NITZAN, DORIT;ETSION, IZHAK;AND OTHERS;SIGNING DATES FROM 20090527 TO 20090804;REEL/FRAME:024081/0826 Owner name: YISSUM RESEARCH DEVELOPMENT COMPANY OF THE HEBREW Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:BARENHOLZ, YECHEZKEL;NITZAN, DORIT;ETSION, IZHAK;AND OTHERS;SIGNING DATES FROM 20090527 TO 20090804;REEL/FRAME:024081/0826 |
|
AS | Assignment |
Owner name: TECHNION RESEARCH AND DEVELOPMENT FOUNDATION LTD., Free format text: RE-RECORD TO CORRECT THE ADDRESS OF THE ASSIGNEE ON AN ASSIGNMENT DOCUMENT RECORDED ON MARCH 15, 2010, PREVIOUSLY RECORDED ON REEL 024081 FRAME 0826;ASSIGNORS:BARENHOLZ, YECHEZKEL;NITZAN, DORIT;ETSION, IZHAK;AND OTHERS;SIGNING DATES FROM 20090527 TO 20090804;REEL/FRAME:024155/0172 Owner name: HADASIT MEDICAL RESEARCH SERVICES & DEVELOPMENT LI Free format text: RE-RECORD TO CORRECT THE ADDRESS OF THE ASSIGNEE ON AN ASSIGNMENT DOCUMENT RECORDED ON MARCH 15, 2010, PREVIOUSLY RECORDED ON REEL 024081 FRAME 0826;ASSIGNORS:BARENHOLZ, YECHEZKEL;NITZAN, DORIT;ETSION, IZHAK;AND OTHERS;SIGNING DATES FROM 20090527 TO 20090804;REEL/FRAME:024155/0172 Owner name: YISSUM RESEARCH DEVELOPMENT COMPANY OF THE HEBREW Free format text: RE-RECORD TO CORRECT THE ADDRESS OF THE ASSIGNEE ON AN ASSIGNMENT DOCUMENT RECORDED ON MARCH 15, 2010, PREVIOUSLY RECORDED ON REEL 024081 FRAME 0826;ASSIGNORS:BARENHOLZ, YECHEZKEL;NITZAN, DORIT;ETSION, IZHAK;AND OTHERS;SIGNING DATES FROM 20090527 TO 20090804;REEL/FRAME:024155/0172 |
|
STCF | Information on status: patent grant |
Free format text: PATENTED CASE |
|
CC | Certificate of correction | ||
MAFP | Maintenance fee payment |
Free format text: PAYMENT OF MAINTENANCE FEE, 4TH YR, SMALL ENTITY (ORIGINAL EVENT CODE: M2551) Year of fee payment: 4 |
|
MAFP | Maintenance fee payment |
Free format text: PAYMENT OF MAINTENANCE FEE, 8TH YR, SMALL ENTITY (ORIGINAL EVENT CODE: M2552); ENTITY STATUS OF PATENT OWNER: SMALL ENTITY Year of fee payment: 8 |